EAB10003 Erlotinib Hydrochloride in Plasma — SLE

Experimental Details

Sample Preparation

SLE plate procedure

(1) Put the Cleanert SLE plate and 96 well collection plate on the 96-well vacuum manifold

(2) Mix 100 μL of plasma, 10 μL of erlotinib hydrochloride standard aq, 10 μL internal standard solution and 100 μL 10% ammonia together, load the mixture into cartridge

(3) Stand for 5min

(4) Elute with 1400 μL of Methyl-t-butyl ether by gravity, dry with N2, resolve the eluate with 200 μL mobile phase;

LLE procedure

Mix 100 μL of plasma, 10 μL oferlotinib hydrochloride standard aq, 10 μL internal standard solution and 100 μL 10% ammonia together, add the solution into centrifuge tube, then add 1.5mL of Methyl-t-butyl ether into the tube for extraction. The layers were left to separate and the organic aliquot removed, dry with N2, resolve the eluate with 200 μL mobile phase;

Instrumentation

Instrumentation: LC-MS/MS, API 4000

Column: Venusil® Silica, 3×50 mm, 3 μm

Column temperature: 25℃

Mobile phase: 80% ACN aq with 0.1% formic acid, 20% water with 0.1% formic acid

Flow rate: 1 mL/min

Sample Injection: 5 μL

Ion source: ESI – Positive

Scan mode: MRM

 Table 1 MS/MS transitions information of Erlotinib hydrochloride and Erlotinib hydrochloride-d6

Results

(1) Recovery

Same quantity of Methyl-t-butyl is used in the experiment. The follow result shows that the recovery of SLE is higher than that of LLE.

Figure 2 The comparison of recovery between SLE and LLE

(2) Phospholipids Removal

The abundance of phospholipids (496.0/184.0) in plasma sample is detected with LC-MS method. Sample processed with SLE contain 1/100 of phospholipids compared with that processed with LLE.

Figure 3 Peak of drug (A) and phospholipids (B) by SLE method

Figure 4 Peak of drug (A) and phospholipids (B) by SPE method

Ordering Information

 Link : http://t.coralcodes.com:829/application/detail/225